Glycolytic flux-signaling controls mouse embryo mesoderm development

Abstract

How cellular metabolic state impacts cellular programs is a fundamental, unresolved question. Here we investigated how glycolytic flux impacts embryonic development, using presomitic mesoderm (PSM) patterning as the experimental model. First, we identified fructose 1,6-bisphosphate (FBP) as an in vivo sentinel metabolite that mirrors glycolytic flux within PSM cells of post-implantation mouse embryos. We found that medium-supplementation with FBP, but not with other glycolytic metabolites, such as fructose 6-phosphate and 3-phosphoglycerate, impaired mesoderm segmentation. To genetically manipulate glycolytic flux and FBP levels, we generated a mouse model enabling the conditional overexpression of dominant active, cytoplasmic Pfkfb3 (cytoPfkfb3). Overexpression of cytoPfkfb3 indeed led to increased glycolytic flux/FBP levels and caused an impairment of mesoderm segmentation, paralleled by the downregulation of Wnt-signaling, reminiscent of the effects seen upon FBP-supplementation. To probe for mechanisms underlying glycolytic flux-signaling, we performed subcellular proteome analysis and revealed that cytoPfkfb3 overexpression altered subcellular localization of certain proteins, including glycolytic enzymes, in PSM cells. Specifically, we revealed that FBP supplementation caused depletion of Pfkl and Aldoa from the nuclear-soluble fraction. Combined, we propose that FBP functions as a flux-signaling metabolite connecting glycolysis and PSM patterning, potentially through modulating subcellular protein localization.

Hypoxia induces an early primitive streak signature, enhancing spontaneous elongation and lineage representation in gastruloids

“…In particular, the increased amount of NMPs and (pre-)somitic mesodermal cells in NH +Chi compared with conventional NN +Chi gastruloids could be related to glycolysis shaping the neuro-mesodermal lineage choice via FGF/WNT pathway activity during axial elongation ( Bulusu et al, 2017 ; Oginuma et al, 2017 ). Interestingly, the directionality of the effect remains unsettled, with some studies reporting positive regulation of WNT signaling by glycolysis and other studies demonstrating the contrary ( Miyazawa et al, 2021 preprint; Oginuma et al, 2017 , 2020 ). Overall, even though the impact of hypoxia at pre-implantation stages remains to be fully resolved, oxygen concentration and metabolic status appear to be stage- and tissue-specific regulators of developmental gene expression and morphology in ESCs and 3D gastruloids.…”